Paris/July 12

From 2007.igem.org

(Difference between revisions)
(Culture of FtsZ TS)
(Culture of FtsZ TS)
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From the plate of FtsZ TS84 121: <br>
From the plate of FtsZ TS84 121: <br>
isolation of 6 clones of the 121.1 on LBA+tet incubated at 42°C + Culture ON at 30°C
isolation of 6 clones of the 121.1 on LBA+tet incubated at 42°C + Culture ON at 30°C
 +
If one clone do not grow at 42, we'll use it to do the transduction

Revision as of 14:26, 12 July 2007


Preparation of a stock of phage on w121:

David B:

  • Filtration of the Cacl2 and MgSO4 stocks
  • Launch a culture:
    • 15ml LB
    • 225µl Cacl2
    • 450µl MgSO4
    • 90µl DAP
    • 150µl W121 from an ON culture
  • Let grow until OD=0.2 (~1h)
  • Add 100µl P1 (stock 10^10/ml)
  • Incubate 2h
  • Add 200µl Chloroform
  • Vortex, centrifuge (15min @ 3000rpm)
  • Recover the supernatant + add 200µl Chloroform = Stock keep at 4°C

Nicolas C.

Titration of bacteriophages P1

We measure the strength of former P1 preparation :

  • The stock prepared the 3.7.7
  • The stock prepared the 8.7.7

See protocols for details.

Culture of FtsZ TS

We want to isolate a good clone of the strain FstZ TS
From the plate of FtsZ TS84 121:
isolation of 6 clones of the 121.1 on LBA+tet incubated at 42°C + Culture ON at 30°C If one clone do not grow at 42, we'll use it to do the transduction